1. Field of the Invention
This invention relates to a diagnostic method for the detection and measurement of viral specific immunoglobulins in animals or humans utilizing relatively large diametered solid-phase carrier units or balls as the basic unit upon which the immunologic reactions occur.
2. Prior Art
The use of solid phase carriers for detection and measurement of viral specific immunoglobulins is well known. Thus, large diametered (e.g. 1/4" diameter) polystyrene balls have been used as a carrier for purified viral antigens following which viral specific immunoglobulins are attached to the purified antigen and detected by RIA or other tracer methods. Such techniques are set forth in detail in the following prior art:
(1) K. Kalimo, Solid-Phase RIA of Rubella Virus IgG and IgM Antibodies--J. Clin. Microbiol. (August, 1976), pages 117-123;
(2) K. Kalimo, Solid-Phase RIA of Human IgM and IgG Antibodies Against Herpes etc.--Infec. Immunity, (March, 1977), pages 883-889;
(3) P. Arstila, A Solid-Phase RIA for IgG and IgM Antibodies Against Measles Virus--J. Gen. Virol., (1977), pages 34, 167-176;
(4) K. Kalimo, Solid-Phase RIA of Herpes Simplex IgG and IgM Antibodies--J. Immunol. Methods, 14 (1977), pages 183-195; and
(5) K. Kalimo, Solid-Phase RIA of Antiviral IgG and IgM Class Antibodies (Thesis, 1977) U. of Turku--Dept. of Virology--SF 20520--Turku, Finland.
There are substantial drawbacks, however, in making purified viral antigens; and furthermore generally only a single purified antigen is absorbed or coated onto the ball. In contrast, the process of this invention utilizes large-diametered plastic balls of the type heretofore employed, but utilizes them, in a different fashion, to permit the expression of most, if not all, of the antigens of the particular virus on a single large-diametered ball.
Large diameter polystyrene balls have also been coated with an antibody for detection of a specific antigen (AUSRIA II-125.RTM., Abbott Laboratories, North Chicago, Ill.). The AUSRIA methodology does not contemplate the expression of unpurified viral antigen(s) on the polystyrene ball.
The prior art also teaches the production and use of unpurified viral antigens on microcellular structures for the purpose of vaccine production and the like (Nature, Vol. 216 (Oct. 7, 1967), A. L. Van Wezel; and Technical Information, "Microcarrier Cell Culture with Cytodex.TM. 1",Pharmacia Fine Chemicals (Piscataway, N.J.) ). The use of microcarriers carrying unpurified viral antigens for the detection of specific immunoglobulins has substantial drawbacks as well. The process of measurement is time-consuming in that a great number of washing and centrifuging steps must be performed in order to retain the microcarriers and their absorbed specific immunoglobulins for measurement. And because of the washing steps required, one also does lose a significant amount of microcarriers, and their absorbed viral antigen, thus reducing the viral antigen concentration perhaps by a significant amount.
In short, solid phase detection and measurement of viral specific immunoglobulins has been hampered by lack of simple techniques. In the closest prior art known of (Kalimo, supra) viral antigen concentration and purification are major difficult and time-consuming steps. Ihe solid phase assay described herein offers a simple method of viral antigen preparation for specific immunoglobulins detection with advantages: easy preparation, storage and handling, increased sensivity of assays due to increased concentration of stable antigens, and the potential of quantitative assessment of specific antibody response in recent primary infection of pregnant women, newborns, vaccinees, etc.